Cell-free apoptosis in Xenopus egg extracts: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria

Cell. 1994 Oct 21;79(2):353-64. doi: 10.1016/0092-8674(94)90203-8.

Abstract

Apoptotic cell death involves a ritual series of morphological changes, presumably reflecting a conserved molecular pathway. We now report that the nuclear events typical of apoptosis can be reproduced in "apoptotic" Xenopus egg extracts. In this cell-free system, nuclear assembly and protein import are initially normal; after 2-4 hr, however, a process of nuclear destruction ensues involving chromatin condensation and the shrinkage and fragmentation of the nuclei. This apoptotic process, which also occurs in nuclei added exogenously, is blocked by the addition of baculovirus-expressed Bcl-2 protein. To block the disintegration of nuclei that are added later, Bcl-2 must be present during this latent period. "Apoptosis" in these extracts requires a dense organelle fraction enriched in mitochondria. The cell-free system described here provides a novel tool for understanding intracellular events in apoptosis and the inhibitory function of Bcl-2.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Apoptosis*
  • CDC2 Protein Kinase / metabolism
  • Cell Nucleus / ultrastructure
  • Cell-Free System
  • Chromatin / ultrastructure
  • DNA Damage
  • In Vitro Techniques
  • Liver / ultrastructure
  • Ovum / cytology
  • Proto-Oncogene Proteins / physiology*
  • Proto-Oncogene Proteins c-bcl-2
  • Rats
  • Signal Transduction
  • Subcellular Fractions / physiology
  • Xenopus laevis

Substances

  • Chromatin
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • CDC2 Protein Kinase